天津医药

天津医药

• 细胞与分子生物学 •    下一篇

MCP-1转染人脐静脉内皮细胞过表达\沉默后相关信号通路研究

宋恩,李光第,周如丹,赵学凌   

  1. 昆明医科大学第一附属医院骨科
  • 收稿日期:2014-05-26 修回日期:2014-08-07 出版日期:2014-11-15 发布日期:2014-11-15
  • 通讯作者: 赵学凌

The Study of Signaling Pathways in MCP-1Over-Expression / Interference of HUVECs

SONG En ,LI Guang di,ZHOU Ru dan,ZHAO Xue ling   

  1. Orthopedics Department of the First Affiliated Hospital of Kunming Medical University
  • Received:2014-05-26 Revised:2014-08-07 Published:2014-11-15 Online:2014-11-15
  • Contact: ZHAO Xue ling

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摘要：

【摘要】  目的  探讨单核细胞趋化蛋白-1（MCP-1）转染人脐静脉内皮细胞过表达/沉默后相关信号通路与深静脉血栓形成的关系。  方法  培养人脐静脉内皮细胞行免疫荧光、免疫共沉淀检测，构建人MCP-1细胞系过表达/沉默载体，基因表达谱芯片检测人MCP-1细胞系过表达/沉默后转录谱变化并行生物信息技术分析。  结果  培养人脐静脉内皮细胞；构建人MCP-1过表达/干扰载体MCP-1-pCDH-GFP/ MCP-1-LMP shRNAmir1经病毒转染后感染HUVECs；基因芯片检测发现与正常细胞相比，过表达载体MCP-1-pCDH-GFP转染细胞有18条信号通路下调，7条信号通路上调；干扰载体MCP-1-LMP shRNAmir1转染细胞有60条信号通路下调，15条信号通路上调。  结论  MCP-1转染人脐静脉内皮细胞后相关信号通路为深静脉血栓疾病分子层面研究提供新的思路，MCP-1可能在深静脉血栓形成发生发展过程中发挥重要作用。

关键词: 单核细胞, 趋化因子类, 内皮,血管, 脐静脉, 信号传导, 静脉血栓形成, 单核细胞趋化蛋白-1, 人脐静脉内皮细胞

Abstract:

[Abstract]   Objective   To investigate the association between the signaling pathways of MCP-1-pCDH-GFP-trans?
fected cells and deep venous thrombosis (DVT).   Methods   The cultured human umbilical vein endothelial cells (HUVECs)were tested by immunofluorescence and co-immunoprecipitation methods. The constructed MCP-1over-expression/interference vector, and the change of transcription profile were detected by microarray assay and biological information technology analysis.  Results   MCP-1over-expression/interference vector MCP-1-pCDH-GFP/MCP-1-LMP shRNAmir1was constructed and HUVECs were transfected. According to the microarray analysis we found that there were18down-expressed signaling pathways and7up-expressed signaling pathways in MCP-1-pCDH-GFP-transfected cells. There were60downexpressed signaling pathways and15up-expressed signaling pathways in the MCP-1-LMP shRNAmir1transfected cells.   Conclusion   Signaling pathways of MCP-1plays an important role in DVT formation.which may provide us a new way to study molecular mechanism of DVT.

Key words: monocytes, chemotactic factors, endothelium,vascular, umbilical veins, signal transduction, venous thrombosis, monocyte chemoattractant protein-1, human umbilical vein endothelial cells